Xinghong Yang

Overexpression of chloroplastic monodehydroascorbate reductase enhanced tolerance to temperature and methyl viologen-mediated oxidative stresses.

A tomato (Lycopersicon esculentum Mill.) monodehydroascorbate reductase gene (LeMDAR) was isolated. The LeMDAR-green fluorescence protein (GFP) fusion protein was targeted to chloroplast in Arabidopsis mesophyll protoplast. RNA and protein gel blot analyses confirmed that the sense- and antisense- LeMDAR were integrated into the tomato genome. The MDAR activities and the levels of reduced ascorbate (AsA) were markedly increased in sense transgenic lines and decreased in antisense transgenic lines compared with wild-type (WT) plants. Under low and high temperature stresses, the sense transgenic plants showed lower level of hydrogen peroxide (H(2)O(2)), lower thiobarbituric acid reactive substance (TBARS) content, higher net photosynthetic rate (P(n)), higher maximal photochemical efficiency of PSII (F(v)/F(m)) and fresh weight compared with WT plants. The oxidizable P700 decreased more obviously in WT and antisense plants than that in sense plants at chilling temperature under low irradiance. Furthermore, the sense transgenic plants exhibited significantly lower H(2)O(2) level, higher ascorbate peroxidase (APX) activity, greater P(n) and F(v)/F(m) under methyl viologen (MV)-mediated oxidative stresses. These results indicated that overexpression of chloroplastic MDAR played an important role in alleviating photoinhibition of PSI and PSII and enhancing the tolerance to various abiotic stresses by elevating AsA level.

Glycine betaine‐mediated potentiation of HSP gene expression involves calcium signaling pathways in tobacco exposed to NaCl stress

Glycine betaine (GB) can enhance heat tolerance and the accumulation of heat-shock protein (HSP) in plants, but the effects of GB on HSP accumulation during salt stress were not previously known. To investigate the mechanism of how GB influences the expression of HSP, wild-type tobacco (Nicotiana tabacum) seedlings pretreated with exogenous GB and BADH-transgenic tobacco plants that accumulated GB in vivo were studied during NaCl stress. A transient Ca(2+) efflux was observed in the epidermal cells of the elongation zone of tobacco roots after NaCl treatment for 1-2 min. After 24 h of NaCl treatment, an influx of Ca(2+) was observed; a low concentration of GB significantly increased NaCl-induced Ca(2+) influx. GB increased the intracellular free calcium ion concentration and enhanced the expression of the calmodulin (CaM) and heat-shock transcription factor (HSF) genes resulting in potentiated levels of HSPs. Pharmacological experiments confirmed that Ca(2+) and CaM increased HSFs and HSPs gene expression, which coincided with increased the levels of HSP70 accumulation. These results suggest a mechanism by which GB acted as a cofactor in the NaCl induction of a Ca(2+) -permeable current. A possible regulatory model of Ca(2+) -CaM in the signal transduction pathway for induction of transcription and translation of the active HSPs is described.

Photosynthesis, sucrose metabolism, and starch accumulation in two NILs of winter wheat

The photosynthetic oxygen evolution rate, Hill reaction activity of seedlings and photosynthetic parameter, Pn–Ci curve and some source-sink metabolism-related enzyme activities, and substance content of flag leaves were measured by using two wheat near isogenic lines with significant differences in the photosynthetic rate of the 154 (high photosynthetic rate) and 212 (low photosynthetic rate) lines as materials. The results showed that the maximal carboxylation efficiency (Vcmax) and Hill reaction activity were higher in line 154 than that of line 212. The Pn in flag leaves of line 154 was significantly higher than that of line 212 during the anthesis to grain-filling stage. Higher leaf sucrose phosphate synthase activity, grain sucrose synthase activity, and grain ADPG pyrophosphorylase activity ensured that the photosynthate of line 154 could be transported to grains and translated into starch in a timely and effective manner, which also contributed to the maintenance of its high photosynthetic rate. Eventually, all of these factors of line 154 resulted in its higher grain yield compared with the low photosynthetic rate of line 212.

Constitutive expression of CaHSP22.5 enhances chilling tolerance in transgenic tobacco by promoting the activity of antioxidative enzymes

Chilling stress limits the productivity and geographical distribution of many organisms throughout the world. In plants, the small heat shock proteins (sHSPs) belong to a group of proteins known as chaperones. The sweet pepper (Capsicum annuum L.) cDNA clone CaHSP22.5, which encodes an endoplasmic reticulum-located sHSP (ER-sHSP), was isolated and introduced into tobacco (Nicotiana tabacum L.) plants and Escherichia coli. The performance index and the maximal efficiency of PSII photochemistry (Fv/Fm) were higher and the accumulation of H2O2 and superoxide radicals (O2-) was lower in the transgenic lines than in the untransformed plants under chilling stress, which suggested that CaHSP22.5 accumulation enhanced photochemical activity and oxidation resistance. However, purified CaHSP22.5 could not directly reduce the contents of H2O2 and O2- in vitro. Additionally, heterologously expressed recombinant CaHSP22.5 enhanced E. coli viability under oxidative stress, helping to elucidate the cellular antioxidant function of CaHSP22.5 in vivo. At the same time, antioxidant enzyme activity was higher, which was consistent with the lower relative electrolyte conductivity and malondialdehyde contents of the transgenic lines compared with the wild-type. Furthermore, constitutive expression of CaHSP22.5 decreased the expression of other endoplasmic reticulum molecular chaperones, which indicated that the constitutive expression of ER-sHSP alleviated endoplasmic reticulum stress caused by chilling stress in plants. We hypothesise that CaHSP22.5 stabilises unfolded proteins as a chaperone and increases the activity of reactive oxygen species-scavenging enzymes to avoid oxidation damage under chilling stress, thereby suggesting that CaHSP22.5 could be useful for improving the tolerance of chilling-sensitive plant types.

Ectopic expression of StCBF1 and ScCBF1 have different functions in response to freezing and drought stresses in Arabidopsis

Solanum tuberosum potato species constitute the bulk of economically and agronomically important potato production. However, S. tuberosum is a drought- and frost-sensitive species that is incapable of acclimating to the cold. Solanum commersonii is a tuber-bearing wild potato species that exhibits greater frost and drought resistance than S. tuberosum. CBF/DREB (C-REPET BINDING FACTOR/DROUGHT RESPONSE ELEMENT BINGING FACTOR) transcription factors play important roles in response to a variety of abiotic stresses, such as cold, drought and salt stresses. To explore different functions between S. tuberosum CBF1 (StCBF1) and S. commersonii CBF1 (ScCBF1), Arabidopsis was transformed with the ScCBF1 and StCBF1 genes driven by a constitutive CaMV35S promoter. Our results reveal that the ScCBF1 transgenic lines are much more tolerant to freezing and drought than the StCBF1 transgenic lines. The development of transgenic plants was altered, resulting in dwarf phenotype with delayed flowering and thicker and additional rosette leaves. The expression levels of several COR (COLD-RESPONSIVE) genes and development-related genes, including genes that inhibited plant growth (GA2ox7, RGL3) and delayed flowering (FLC) were higher in transgenic plants. These results suggest that these two potato CBF1 play important roles in the plant response to abiotic stress and can influence plant growth and development, and ScCBF1 plays a more pronounced function than StCBF1.

Stimulation of cyclic electron flow around PSI as a response to the combined stress of high light and high temperature in grape leaves

Changes in cyclic electron flow (CEF) around PSI activity after exposing grape (Vitis vinifera L.) seedling leaves to the combined stress of high temperature (HT) and high light (HL) were investigated. The PSII potential quantum efficiency (Fv/Fm) decreased significantly under exposure to HT, and this decrease was greater when HT was combined with HL, whereas the PSI activity maintained stable. HT enhanced CEF mediated by NAD(P)H dehydrogenase remarkably. Compared with the control leaves, the half-time of P700+ re-reduction decreased during the HT treatment; this decrease was even more pronounced under the combined stress, implying significantly enhanced CEF as a result of the treatment. However, the heat-induced increase in nonphotochemical quenching (NPQ) was greater under HL, accompanied by a greater enhancement in high-energy state quenching. These results suggest that the combined stress of HT and HL resulted in severe PSII photoinhibition, whereas CEF showed plasticity in its response to environmental stress and played an important role in PSII and PSI photoprotection through accelerating generation of the thylakoid proton gradient and the induction of NPQ.